Agarose gel electrophoresis
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General Information
General Procedure
- Cast a gel
- Place it in gel box in running buffer
- Load samples
- Run the gel
- Image the gel
Buffers
- TAE - better resolution of fragments >4kb;
- TBE - better resolution of 0.1-3kb fragments; TBE is better suited for high-voltage (>150V) electrophoresis than TAE because of its higher buffering capacity and lower conductivity;
Loading dyes
dye | 0.5-1.5% agarose | 2.0-3.0% agarose* |
xylene cyanol | 4000-5000 bp | 750 bp |
bromophenol blue | 400-500 bp | 100 bp |
*sieving agarose
Specific Protocols
Knight:Agarose gel electrophoresis
Notes
- If you are getting unexpected bands on your gel you may want to look at the common issues in agarose gel electrophoresis page.
- If you have no experience with gel electrophoresis or are explaining it to someone new, here is a cute java demo of what happens.